Method of Treating Inflammation

ABSTRACT

The present invention concerns methods of treating systemic, regional, or local inflammation from a patient suffering or at risk of inflammation comprising administration of a therapeutically effective dose of a sorbent that sorbs an inflammatory mediator in said patient. In some preferred embodiments, the sorbent is a biocompatible organic polymer.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No.15/172,209, filed Jun. 3, 2016, which is a continuation of U.S. patentapplication Ser. No. 13/637,775 filed on Mar. 8, 2013, which is theNational Stage of International Application No. PCT/US2011/030919, filedApr. 1, 2011, which claims the benefit of U.S. Provisional ApplicationNo. 61/319,993, filed Apr. 1, 2010, the disclosures of which areincorporated herein by reference in their entireties.

TECHNICAL FIELD

The invention concerns, inter alia., methods of treating systemic,regional, or local inflammation from a patient suffering or at risk ofsuch inflammation.

BACKGROUND

An inflammatory response occurs in animals when cells or tissues areinjured by bacteria, trauma, toxins, heat, or other agents, which can becollectively referred to as “Inflammatory Agents.” The nature andcharacter of a given inflammatory response is regulated by the complexinteraction of a variety of pro-inflammatory or anti-inflammatorystimulators or mediators, which are synthesized and released by cellsand tissues. Some known species of pro-inflammatory or anti-inflammatorystimulators or mediators include cytokines, nitric oxide, thromboxanes,leukotrienes, phospholipids like platelet-activating factor,prostaglandins, kinins, complement factors, coagulation factors,superantigens, monokines, chemokines, interferons, free radicals,proteases, arachidonic acid metabolites, prostacyclins, beta endorphins,myocardial depressant factors, anandamide, 2-arachidonoylglycerol,tetrahydrobiopterin, cell fragments and chemicals including histamine,bradykinin, and serotonin. The discovery of new (i.e., previouslyunrecognized) species of pro-inflammatory or anti-inflammatorystimulators or mediators is an ongoing process.

The nature and intensity of inflammatory responses differ, depending onthe site which has been invaded, and on the character of theInflammatory Agent(s), and the interaction of pro-inflammatory oranti-inflammatory stimulators or mediators involved. The inflammatoryresponse, when regulated and localized, is beneficial. If not regulatedand generalized, however, the inflammatory response can causesignificant tissue injury and even death.

Cytokines are one class of proteins produced predominantly bymacrophages, monocytes, neutrophils and lymphocytes typically inresponse to a viral, bacterial, fungal or parasitic infection, as wellas in response to T cell stimulation during an immune response.Cytokines are known to be synthesized by other cell types, such asstromal cells like fibroblasts, endothelial cells and smooth musclecells, as well as epithelial cells, keratinocytes and hepatocytes.Cytokines are normally present in very low concentrations in the bloodor tissues.

The structures and activities of cytokines have been the subject of manystudies. It has become apparent that cytokines possess a wide spectrumof immunological and non-immunological activities. Cytokines affectdiverse physiologic functions, such as cell growth, differentiation,homeostasis and pathological physiology. The art shows that cytokineshave multiple biological activities and interact with more than one celltype. Cytokines are also known to be capable of stimulating their ownsynthesis, as well as the production of other cytokines from a varietyof cell types. This phenomenon is called the “cytokine cascade.”Cytokine cascades are often associated with systemic changes arisingfrom infection and tissue injury and, in this context, they serve amyriad of biological functions. For example, various cytokines,categorized as the interleukins (IL), interferons (IF), and tumornecrosis factor (TNF), are produced during immune and inflammatoryresponses. These cytokines beneficially control various aspects of theseresponses. In this situation, the cytokine cascade mediates normal hostdefense responses, cell regulation, and cell differentiation.

Under cascade circumstances, the function of cytokine production canbecome disordered. This disorder can lead to the presence of larger thannormal concentrations of cytokines. When the cytokine cascade becomesdisordered, there can be a rapid extension and amplification of theintended localized host response in such a way that only one or a fewinitiating stimuli trigger the eventual release and participation ofscores of host mediators. Although a number of features of the hostresponse assist in fighting off invasion, an overly robust or poorlymodulated endogenous response can rapidly accelerate to produce otherprofound alterations in host homeostasis at the cellular, tissue, andsystemic levels. As a result, cytokine expression in a region of thebody where tissues or organs are legitimately subject to bacterialinfection or an immune response challenge, can, when disordered, lead tounwanted destruction of healthy tissue elsewhere in the body. Largerthan normal concentrations of certain cytokines can cause disease andother deleterious health effects, some of which can be lethal.

A disordered cytokine cascade that leads to the increased presence ofthe cytokines IL-1 and TNF can, alone or in combination, cause a statein animals clinically identical to systemic inflammatory responsesyndrome (SIRS), sepsis, and more severe variants of sepsis calledsevere sepsis (sepsis with organ dysfunction) and septic shock (severesepsis with refractory hypotension). These conditions can arise due tothe individual, combined, and concerted effects of a large number ofcytokines. Severe sepsis and septic shock afflicts more than 750,000Americans every year. Cytokine-induced sepsis can be brought about byinfection by a variety of microorganisms, including not only bacteriabut also viruses, fungi, and parasites. SIRS or sepsis can also beinitiated by host response to invasion in general, such as by cancer oras a result of major surgery or trauma. Septic shock is a potentiallylethal cytokine-mediated clinical complication against which there is nogenerally effective therapeutic approach.

One of the best studied examples of cytokine-induced septic shock is thecase of infection by gram-negative bacteria. The appearance of bacterialendotoxins, such as lipopolysaccharide (LPS), in the host bloodstream isbelieved to lead to the endogenous production of a variety of hostfactors that directly and indirectly mediate the toxicity of LPS. Thesehost-derived mediators include many now well-recognized inflammatorycytokines, as well as endocrine hormones, in addition to a number ofother endogenous factors such as leukotrienes and platelet activatingfactor. Among the interacting factors that together comprise thecytokine cascade, the cytokine TNF-alpha is believed to be the mostimportant identified to date. During the ensuing cytokine cascade, themediators that appear early in the invaded host are thought to triggerthe release of later appearing factors. Many of the cytokine mediatorsnot only exert direct functions at the targeted tissues, but also atother local and remote tissues, where subsequent responses to othermediators produced during the cascade occur, and so on. The result, ifunchecked, can be a multifaceted pathological condition, which ischaracterized most prominently by deleterious hemodynamic changes, organdysfunction and coagulopathy leading to multiple organ failure and,often, to death.

Multiple attempts have been made and still many others are currentlyunderway to block specific mediators of this response. These attempts,however, have been relatively unsuccessful. Therapy aimed at singlemediators cannot effectively attenuate the entire response. Furthermore,it is both the duration and intensity of inflammation that correlatesbest with outcome. Generally, higher concentrations of cytokines andlonger duration of over-expression of proinflammatory cytokines areassociated with higher mortality. Systemic inflammation results in organinjury which results in the prolongation of the inflammatory responseand thus, more organ injury.

Similarly profound, but often less lethal, physiologic effects can occuras a result of abnormal production of certain cytokines, without thepresence of exogenous bacterial toxins. As one example, cytokineTNF-alpha has been found to be an anti-tumor cytokine. As a result,TNF-alpha has been expected to be useful as an antitumor agent. However,it has been discovered that TNF-alpha is identical with cachectin, whichis a cachexia-inducing factor. The disordered production of TNF-alphahas also been correlated with, not only severe sepsis and septic shock,but the incidence of rheumatoid arthritis, adult respiratory distresssyndrome (ARDS), the severity of viral hepatitis, myocardial ischemia,and the inhibition of myocardial contraction. Also, TNF has recentlybeen shown to be involved in initiating the expression of humanimmunodeficiency virus in human cells that carry latent virus, whichcould be a contributing factor in the expression of latent AIDS virus incertain individuals. Furthermore, a correlation between the TNF level inthe blood and blood pressure has also been observed. As TNF levelsincrease, blood pressure decreases, which can lead to seriouscomplications such as kidney failure. Hypotension and in severe cases,hemodynamic collapse or shock, can be caused by cytokines such as TNFthrough endothelial damage, leading to loss of fluid from theintravascular space to the surrounding tissues, as well as through TNFand other cytokine stimulation of inducible nitric oxide synthase, thatleads to myocardial depression and peripheral vasodilation.

TNF-alpha has been observed to stimulate production of other types ofcytokines, such as IL-1, etc. Cytokine IL-1 is known to be an importantagent for inducing and transmitting the systemic biological responseagainst infection and inflammation. IL-1 induces the usual, desirableresponses observed in inflammation in general, such as fever, increaseof leukocytes, activation of lymphocytes, and induction of biosynthesisof acute phase protein in liver. This cytokine is known to have a strongantitumor activity.

When IL-1 is produced in abnormally larger amounts, however, the resultmay contribute to the severity of chronic inflammatory diseases, such asrheumatoid arthritis. Thus, the abnormal activation of various cytokinessuch as the interleukins (IL) and tumor necrosis factor (TNF) isbelieved responsible for the tissue damage and pain that occurs invarious inflammatory conditions like rheumatoid arthritis. In rheumatoidarthritis, levels of TNF, IL-1, IL-6 and IL-8 increase dramatically andcan be detected in the synovial fluid. The cytokine cascade induced byexpression of these cytokines results in depressed lipoproteinmetabolism as well as bone and cartilage destruction.

As another example, the cytokine IL-6 plays an important role inantibody production in B cells. The cytokine IL-6 also is an importantfactor in body systems, e.g., the hematopoietic system, nervous system,and the liver, as well as in immune system. For example, IL-6 iseffective for inducing proliferation and differentiation of T cells,inducing the production of protein at acute phase by acting on hepaticcells, and promoting the growth of cells in bone marrow.

A correlation between the abnormal secretion of IL-6 and various diseasestates (e.g., autoimmune diseases, such as hypergammaglobulinemia,chronic articular rheumatism, and systemic lupus erythematosus; theabnormal state of polyclonal B cells, as well as in the development ofthe abnormal state of monoclonal B cells such as myeloma cells;Castleman's disease accompanied with tumor of the lymph nodes, for whichthe cause is unknown; primary glomerular nephritis; and the growth ofmesangial cells) has been observed.

As yet another example, in bacterial infections, cytokines such as IL-8act as a signal that attracts white blood cells such as neutrophils tothe region of cytokine expression. In general, the release of enzymesand superoxide anions by neutrophils is essential for destroying theinfecting bacteria. However, if cytokine expression causes neutrophilsto invade, for example, the lungs, release of neutrophil enzymes andsuperoxide anion can result in the development of adult respiratorydistress syndrome (ARDS), which can be lethal.

Despite their diverse and myriad functions, most cytokines share onecommon feature. Although most cytokines are found in the size andmolecular weight range of 8 to 80 kilodaltons, the majority of cytokinesare within a narrow size and molecular weight range of 8 to 51kilodaltons. This size characteristic is extremely important for theclearance of cytokines from the blood.

In disease states where the kidney has failed—which is often the case inseptic shock—hemodialysis or hemofiltration membranes are used assubstitutes for the glomerular membrane of the kidney. Artificialmembranes, however, are severely limited in their ability to clearcytokines from the blood due to their inadequate porosity. In fact, thepredominant mechanism by which these membranes remove cytokines inclinical practice is not filtration, but rather nonspecific surfaceadsorption (J. Am Soc Nephrol 1999 April; 10(4): 846-53, Cytokineremoval during continuous hemofiltration in septic patients, De Vriese AS, Colardyn F A, Philippe J J, Vanholder R C, De Sutter J H, Lameire NH). Typically these membranes have 0.5 to 2 square meters of surfacearea available for adsorption that becomes saturated within the first 30to 90 minutes of treatment (Biomaterials September 1999; 20(17):1621-34,Adsorption of low molecular weight proteins to hemodialysis membranes:experimental results and simulations, Valette P, Thomas M, Dejardin P).An improved external device taught in the art uses certain sorbentpolymers to externally interact with blood to reduce levels of cytokinesor other species of pro-inflammatory or anti-inflammatory stimulators ormediators in blood or physiologic fluids with significant specificity.See, U.S. Pat. Nos. 7,556,768; 6,416,487; and 5,904,663. While suchmethods are advantageous in treating certain inflammatory conditions,there is a need in the art for a simpler method that does not requireuse of an external removal device.

SUMMARY

In some aspects, the invention concerns methods of treating systemic,regional, or local inflammation from a patient suffering or at risk ofinflammation comprising administration of a therapeutically effectivedose of a sorbent for an inflammatory mediator (for example, a sorbentthat sorbs the mediator) in the patient. In some methods, theinflammatory mediator is one or more of enzymes, cytokines,prostaglandins, eicosanoids, leukotrienes, kinins, complement,coagulation factors, toxins, endotoxins, enterotoxins,lipopolysaccharide, substances that induce apoptosis of cells, causticsubstances, bile salts, fatty acids, phospholipids, oxidized byproducts,reactive oxygen species, oxygen radicals, surfactants, ions, irritantsubstances, cell fragments, interferon and immunomodulatory antibodies,biologics or drugs.

Certain inflammatory mediators are present in physiological fluid or acarrier fluid in the patient. Physiological fluids includenasopharyngeal, oral, esophageal, gastric, pancreatic, hepatic, pleural,pericardial, peritoneal, intestinal, prostatic, seminal, vaginalsecretions, tears, saliva, mucus, bile, blood, lymph, plasma, serum,synovial fluid, cerebrospinal fluid, urine, and interstitial,intracellular, and extracellular fluid.

In some preferred methods, the dose of sorbent is administered orally,via a feeding tube, peritoneally, or rectally.

The inflammatory mediator may be associated with systemic inflammatoryresponse syndrome (SIRS) or sepsis, such as from viral, bacterial,fungal or parasitic infection, autoimmune disease, surgery, cytotoxicchemotherapy, bone marrow manipulation, major tissue injury or trauma,mesenteric hypoperfusion, gut-mucosal injury, malaria, gastrointestinalinflammatory disease, enteric infection, influenza, acute lunginflammation such as acute respiratory distress syndrome or acute lunginjury, pulmonary embolism, pancreatitis, autoimmune and collagenvascular diseases, transfusion-related diseases, burn injury, smoke orinhalation lung injury, graft versus host disease, ischemia orinfarction, reperfusion injury, hemorrhage, anaphylaxis, drug overdose,radiation injury or chemical injury. In some embodiments, theinflammatory mediator results from a disease caused biowarfarepathogens, toxins or agents, such as viral hemorrhagic fevers,hantavirus cardiopulmonary syndrome (hantaviruses), cholera toxin,botulinum toxin, Ricin toxin, Q fever (Coxiella burnetii), Typhus(Rickettsia prowaszekii), or Psittacosis (Chlamydia psittaci).

In certain embodiments of the invention, the sorbent is a biocompatiblepolymer.

Some polymers can be supplied as a slurry, or suspension, or dry powderor other dry particulate capable of being wetted. In some methods, thesorbent is supplied as a slurry or suspension packaged in either singledose or multidose packages for oral administration. In other methods,the sorbent is supplied as a slurry or suspension packaged in eithersingle dose or multidose packages for administration by enema or feedingtube or any combination therein.

The polymer can also be supplied as a dry powder or other dryparticulate capable of being wetted externally or internally in thealimentary canal, including in the gastric or enteric environment, withor without the addition of wetting agents such as ethyl or isopropylalcohol. In yet other embodiments, the polymer is supplied as tablet,dry powder, other dry particulate, capsule or suppository packaged inbottles or blister packs for administration.

In some methods, the polymer materials are not metabolizable by humanand animal.

Some polymers comprise particles having a diameter in the range for 0.1micron meters to 2 centimeters. Certain polymers are in the form ofpowder, beads or other regular or irregularly shaped particulate. Thepore structure of some polymers is such that the total pore volume ofpore size in the range of 50 < to 3000 Å is greater than 0.5 cc/g to 3.0cc/g dry polymer. In some embodiments, the polymer has a pore structuresuch that the total pore volume of pore size in the range of 50 Å to3000 Å is greater than 0.5 cc/g to 3.0 cc/g dry polymer; wherein theratio of pore volume between 50 Å to 3,000 Å (pore diameter) to porevolume between 500 Å to 3,000 Å (pore diameter) of the polymer issmaller than 200:1; and the ratio of pore volume between 50 Å to 3,000 Å(pore diameter) to pore volume between 1,000 Å to 3,000A (pore diameter)of the polymer is greater than 20:1.

In some embodiments, the polymer is a coated polymer comprising at leastone crosslinking agent and at least one dispersing agent. The dispersingagents can be selected from such as hydroxyethyl cellulose, hydroxypopylcellulose, poly(hydroxyethyl methacrylate), poly(hydroxyethyl acrylate),poly(hydroxypropyl methacrylate), poly(hydroxypropyl acrylate),poly(dimethylaminoethyl methacrylate), poly(dimethylaminoethylacrylate), poly(diethylamimoethyl methacrylate), poly(diethylaminoethylacrylate), poly(vinyl alcohol), poly(N-vinylpyrrolidinone), salts ofpoly(methacrylic acid), and salts of poly(acrylic acid) and mixturesthereof; the crosslinking agent selected from a group consisting ofdivinylbenzene, trivinylbenzene, divinylnaphthalene,trivinylcyclohexane, divinylsulfone, trimethylolpropane trimethacrylate,trimethylolpropane dimethacrylate, trimethylolpropane triacrylate,trimethylolpropane diacrylate, pentaerythrital dimethacrylates,pentaerythrital trimethacrylates, pentaerythrital, tetramethacrylates,pentaerythritol diacrylates, pentaerythritol triiacrylates,pentaerythritol tetraacrylates, dipentaerythritol dimethacrylates,dipentaerythritol trimethacrylates, dipentaerythritoltetramethacrylates, dipentaerythritol diacrylates, dipentaerythritoltriacrylates, dipentaerythritol tetraacrylates, divinylformamide andmixtures thereof; and the polymer is developed simultaneously with theformation of the coating, wherein the dispersing agent is chemicallybound to the surface of the polymer.

Some preferred polymers comprise residues from one or more monomersselected from divnylbenzene and ethylvinylbezene, styrene, ethylstyrene,acrylonitrile, butyl methacrylate, octyl methacrylate, butyl acrylate,octyl acrylate, cetyl methacrylate, cetyl acrylate, ethyl methacrylate,ethyl acrylate, vinyltoluene, vinylnaphthalene, vinylbenzyl alcohol,vinylformamide, methyl methacrylate, methyl acrylate, trivinylbenzene,divinylnaphthalene, trivinylcyclohexane, divinylsulfone,trimethylolpropane trimethacrylate, trimethylolpropane dimethacrylate,trimethylolpropane triacrylate, trimethylolpropane diacrylate,pentaerythritol dimethacrylate, pentaerythritol trimethacrylate,pentaerythritol tetramethacrylate, pentaerythritol diacrylate,pentaerythritol triiacrylate, pentaerythritol tetraacrylate,dipentaerythritol dimethacrylate, dipentaerythritol trimethacrylate,dipentaerythritol tetramethacrylate, dipentaerythritol diacrylate,dipentaerythritol triacrylate, dipentaerythritol tetraacrylate,divinylformamide and mixtures thereof.

Some suitable polymer include ion exchange polymers. In someembodiments, the polymer is a cellulosic polymer. In some embodiments,the polymers may be derivatized. Some polymers may be modified with anantibody or ligand. Such polymer may be porous or solid

Certain preferred polymers are porous highly crosslinked styrene ordivinylbenzene copolymer. In some embodiments, the porous highlycrosslinked styrene or divinylbenzene copolymer is a macroporous ormesoporous styrene-divinylbenzene-ethylstyrene copolymer subjected to apartial chloromethylation to a chlorine content of up to 7% molecularweight. In certain embodiments, the porous highly crosslinked styrene ordivinylbenzene copolymer is a hypercrosslinked polystyrene produced fromcrosslinked styrene copolymers by an extensive chloromethylation and asubsequent post-crosslinking by treating with a Friedel-Crafts catalystin a swollen state. In other embodiments, the porous highly crosslinkedstyrene or divinylbenzene copolymer is a hypercrosslinked polystyreneproduced from crosslinked styrene copolymers by an extensive additionalpost-crosslinking in a swollen state with bifunctional crosslinkingagents selected from the group comprising of monochlorodimethyl etherand p-xylilene dichloride.

In another aspect of the invention, the polymer is a hydrophilic selfwetting polymer that can be administered as dry powder or dryparticulate containing hydrophilic functional groups such as chlorine,amines, hydroxyl, sulfonate, and carboxyl groups

Certain polymer may be pyrolyzed.

The invention also concerns methods of preventing inhabitation of normalcell proliferation, activity, growth or regeneration comprisingadministration of a therapeutically effective dose of a sorbent to apatient that sorbs an inflammatory mediator in the patient. In someembodiments, the inhibition concerns cartilage and cartilage cells. Incertain embodiments, the administration is accomplished by temporarilyplacing the sorbent in the joint or articular space.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows a comparison of Botulinum Toxin A removal by porous polymercompared to non porous polymer and vehicle control.

FIG. 2 presents a graph of C-reactive protein (CRP) concentration overtime in animals with cecal ligation puncture induced infection andsepsis receiving an oral dose of a 25% slurry of porous beads in watercompared to control water only.

FIG. 3 shows percent of animals with cecal ligation puncture inducedinfection and sepsis that are living versus time after oraladministration of porous polymer.

FIG. 4 presents a comparison mean clinical score for control vstreatment for TNBS Colitis model.

DETAILED DESCRIPTION OF ILLUSTRATIVE EMBODIMENTS

In some aspects, the invention concerns methods of treating systemic,regional, or local inflammation from a patient suffering or at risk ofinflammation by administering a therapeutically effective dose of asorbent that sorbs an inflammatory mediator in the patient. Aninflammatory mediator as we have defined it is any substance that candirectly or indirectly elicit irritation, inflammation, injury or deathof local or remote cells, tissues or organs. Inflammatory mediatorsinclude, but are not limited to, enzymes, cytokines, prostaglandins,eicosanoids, leukotrienes, kinins, complement, coagulation factors,toxins, endotoxins, enterotoxins, lipopolysaccharide, substances thatinduce apoptosis of cells including proteins such as Fas ligand, cellfragments, caustic substances such as acidic or basic secretions, bilesalts, fatty acids, phospholipids, oxidized byproducts, reactive oxygenspecies, oxygen radicals, surfactants, ions, and irritant chemicals.Inflammatory mediators also include agents administered exogenously suchas interferon and immunomodulatory antibodies, biologics or drugs.Indirect action could include, for instance, initiating a chain ofevents that leads to an activation of the pro-inflammatory immuneresponse.

In some embodiments of the invention, the sorbent is used to sorb aninflammatory mediator present in physiological fluid or a carrier fluidin the patient. Physiologic fluids are liquids that originate from thebody and can include, but are not limited to, nasopharyngeal, oral,esophageal, gastric, pancreatic, hepatic, pleural, pericardial,peritoneal, intestinal, prostatic, seminal, vaginal secretions, as wellas tears, saliva, mucus, bile, blood, lymph, plasma, serum, synovialfluid, cerebrospinal fluid, urine, and interstitial, intracellular, andextracellular fluid, such as fluid that exudes from burns or wounds.Carrier fluids are exogenously administered liquids that include, butare not limited to, liquids administered orally, via a feeding tube,peritoneally, or rectally such as an enema or colonic wash.

The compositions of the present invention are useful in treating a widevariety of inflammatory conditions. For example, the methods can be usedon conditions where the inflammatory mediator is associated withsystemic inflammatory response syndrome (SIRS) or sepsis, caused bysituations such as viral, bacterial, fungal or parasitic infections,autoimmune disease, surgery, cytotoxic chemotherapy, bone marrowmanipulation, major tissue injury or trauma, mesenteric hypoperfusion,gut-mucosal injury, malaria, gastrointestinal inflammatory disease,enteric infection, influenza, acute lung inflammation such as acuterespiratory distress syndrome or acute lung injury, pulmonary embolism,pancreatitis, autoimmune and collagen vascular diseases,transfusion-related diseases, burn injury, smoke and inhalation lunginjury, graft versus host disease, ischemia or infarction, reperfusioninjury, hemorrhage, anaphylaxis, drug overdose, radiation injury andchemical injury. The methods are also useful for treating conditionswhere a disease is caused specifically by potential biowarfarepathogens, toxins or agents including, but not limited to, anthrax(Bacillus anthracis), influenza, smallpox, SARS coronavirus, bubonicplague (Yersinia pestis), viral hemorrhagic fevers (filoviruses likeEbola and Marburg and arenaviruses like Lassa virus), tularemia(Francisella tularensis), hantavirus cardiopulmonary syndrome(hantaviruses), cholera toxin, botulinum toxin, Ricin toxin, Q fever(Coxiella burnetii), Typhus (Rickettsia prowaszekii), and Psittacosis(Chlamydia psittaci).

In addition to treatment of diseases or other inflammatory conditions orinjuries, compositions disclosed herein may also be usefulprophylactically for the prevention of such conditions.

Inflammatory mediators can inhibit normal cell proliferation, activity,growth or regeneration. Cartilage is one such example. It is generallybelieved that cytokines and inflammatory mediators andmetalloproteinases (enzymes) are partly responsible for cartilage deathin joints (rheumatoid arthritis, osteoarthritis, etc). Some believe thattheir presence in synovial fluid prevents cartilage and cartilage cellsfrom being able to regenerate and regrow. By administering thesepolymers so as to place the sorbent (temporarily, in a preferredembodiment) in the joint or articular space (either in a contained oruncontained form) to sorb these mediators, cartilage and cartilage cellscan be allowed to regrow in situ.

The compositions of the instant invention may be administered by methodswell known to those skilled in the art. In some embodiments,administration is topical. Such methods include ophthalmicadministration, administration to skin or wounds, direct administrationinto a body cavity or joint, and delivery to mucous membranes such asnasal, oral, vaginal and rectal delivery or other delivery to thealimentary canal. In some embodiments, such methods include local orsystemic administration through an oral or parenteral route. In someembodiments, the treatment is extracorporeal. Extracorporealadministration would include removal of inflammatory mediators fromblood or physiologic fluids by circulating the fluids through a devicecontaining sorbent and returning it back to the body. Parenteraladministration includes intravenous, intraarterial, subcutaneous,intraperitoneal or intramuscular injection or infusion; or intracranial(including intrathecal or intraventricular, administration).

Pharmaceutical compositions and formulations for topical administrationinclude but are not limited to ointments, lotions, creams, transdermalpatches, gels, drops, suppositories, sprays, liquids and powders.Utilization of conventional pharmaceutical carriers, oily bases,aqueous, powder, thickeners and the like may be used in theformulations.

The pharmaceutical compositions may also be administered in tablets,capsules, gel capsules, slurries, suspensions, and the like.

Penetration enhancers may also be used in the instant pharmaceuticalcompositions. Such enhancers include surfactants, fatty acids, bilesalts, chelating agents, and non-chelating non-surfactants and aregenerally known in the art.

In some preferred embodiments, the administration of sorbent polymer isoral, or rectal, or via a feeding tube or any combination therein.

The polymers of the present invention can be administered once to apatient or in multiple doses.

Polymers useful in the invention may be supplied as a slurry, suspensionor reconstituted from the dry state into a slurry or suspension. In someembodiments, the polymer may be supplied as a slurry or suspensionpackaged in either single dose or multidose bottles for oraladministration. In other embodiments, the polymer may be supplied as aslurry or suspension packaged in either single dose or multidose bottlesfor administration by enema or feeding tube or any combination therein.In certain embodiments, the polymer is supplied as a dry powder capableof being wetted externally or in the alimentary canal with or withoutthe addition of wetting agents such as ethyl alcohol.

The polymer may be supplied as tablet, capsule or suppository packagedin bottles or blister packs for administration. Depending on the use,the polymer may be sterile or non-sterile. The polymer may be sterilizedby standard methods. Such methods are well known to those skilled in theart.

In some embodiments, the polymer materials used as the sorbent aresubstantially not metabolizable by human and animal. Certain polymersmay be irregular or regular shaped particulates such as powders, beads,or other forms with a diameter in the range of 0.1 micron meters to 2centimeters

The polymers used in the instant invention preferably have abiocompatible and hemocompatible exterior surface coatings but are notabsolutely necessary, especially in certain circumstances, such as oralor rectal administration. Certain of these coatings are covalently boundto the polymer particle (beads, for example) by free-radical grafting.The free-radical grafting may occur, for example, during thetransformation of the monomer droplets into polymer beads. Thedispersant coating and stabilizing the monomer droplets becomescovalently bound to the droplet surface as the monomers within thedroplets polymerize and are converted into polymer. Biocompatible andhemocompatible exterior surface coatings can be covalently grafted ontothe preformed polymer beads if the dispersant used in the suspensionpolymerization is not one that imparts biocompatibility orhemocompatibility. Grafting of biocompatible and hemocompatible coatingsonto preformed polymer beads is carried out by activating free-radicalinitiators in the presence of either the monomers or low molecularweight oligomers of the polymers that impart biocompatibility orhemocompatibility to the surface coating.

By “biocompatible”, it is meant that the polymer is capable of contactwith living tissues or organisms without causing harm during the timethat the polymer is in contact with the tissue or organism. In someembodiments, it is intended that the polymer is tolerated by the gut andalimentary canal of the organism. The polymers of the present inventionare preferably non-toxic.

In one embodiment, the present invention relates to a porous polymer forsorbing small to midsize protein molecules and excluding sorption oflarge blood proteins, the polymer comprising a plurality of pores. Thepores sorb small to midsize protein molecules equal to or less than50,000 Daltons.

In some embodiments, the polymer has a preferential pore structure suchthat the total pore volume of pore size in the range of 50 Å to 3000 Åis greater than 0.5 cc/g to 3.0 cc/g dry polymer; wherein the ratio ofpore volume between 50 Å to 3,000 Å (pore diameter) to the pore volumebetween 500 Å to 3,000 Å (pore diameter) of the polymer is smaller than200:1; and the ratio of pore volume between 50 Å to 3,000 Å in diameterto the pore volume between 1,000 Å to 3,000 Å in diameter of the polymeris greater than 20:1. The said ratios can be alternatively specified interms of pore surface area (such as the ratio of pore surface areabetween 50 Å to 3,000 Å to pore surface area between 500 Å to 3,000 Å ofthe polymer); and therefore is an alternative way of specifying the samepore structure.

Some preferred polymers are coated polymers comprising at least onecrosslinking agent and at least one dispersing agent. Suitabledispersing agents include hydroxyethyl cellulose, hydroxypopylcellulose, poly(hydroxyethyl methacrylate), poly(hydroxyethyl acrylate),poly(hydroxypropyl methacrylate), poly(hydroxypropyl acrylate),poly(dimethylaminoethyl methacrylate), poly(dimethylaminoethylacrylate), poly(diethylamimoethyl methacrylate), poly(diethylaminoethylacrylate), poly(vinyl alcohol), poly(N-vinylpyrrolidinone), salts ofpoly(methacrylic acid), and salts of poly(acrylic acid) and mixturesthereof.

Suitable crosslinking agents include divinylbenzene, trivinylbenzene,divinylnaphthalene, trivinylcyclohexane, divinylsulfone,trimethylolpropane trimethacrylate, trimethylolpropane dimethacrylate,trimethylolpropane triacrylate, trimethylolpropane diacrylate,pentaerythrital dimethacrylates, pentaerythrital trimethacrylates,pentaerythrital, tetramethacrylates, pentaerythritol diacrylates,pentaerythritol triiacrylates, pentaerythritol tetraacrylates,dipentaerythritol dimethacrylates, dipentaerythritol trimethacrylates,dipentaerythritol tetramethacrylates, dipentaerythritol diacrylates,dipentaerythritol triacrylates, dipentaerythritol tetraacrylates,divinylformamide and mixtures thereof. Preferably, the polymer isdeveloped simultaneously with the formation of the coating, such thatthe dispersing agent gets chemically bound to the surface of thepolymer.

Preferred polymers include those derived from one or more monomersselected from divnylbenzene and ethylvinylbezene, styrene, ethylstyrene,acrylonitrile, butyl methacrylate, octyl methacrylate, butyl acrylate,octyl acrylate, cetyl methacrylate, cetyl acrylate, ethyl methacrylate,ethyl acrylate, vinyltoluene, vinylnaphthalene, vinylbenzyl alcohol,vinylformamide, methyl methacrylate, methyl acrylate, trivinylbenzene,divinylnaphthalene, trivinylcyclohexane, divinylsulfone,trimethylolpropane trimethacrylate, trimethylolpropane dimethacrylate,trimethylolpropane triacrylate, trimethylolpropane diacrylate,pentaerythritol dimethacrylate, pentaerythritol trimethacrylate,pentaerythritol tetramethacrylate, pentaerythritol diacrylate,pentaerythritol triiacrylate, pentaerythritol tetraacrylate,dipentaerythritol dimethacrylate, dipentaerythritol trimethacrylate,dipentaerythritol tetramethacrylate, dipentaerythritol diacrylate,dipentaerythritol triacrylate, dipentaerythritol tetraacrylate,divinylformamide and mixtures thereof.

Some preferred polymers are ion exchange polymers.

Some preferred polymers are cellulosic polymers. Suitable polymersinclude cross-linked dextran gels such as Sephadex®.

Certain preferred polymers are porous highly crosslinked styrene ordivinylbenzene copolymer. Some of these polymers are a macroporous ormesoporous styrene-divinylbenzene-ethylstyrene copolymer subjected to apartial chloromethylation to a chlorine content of up to 7% molecularweight. Other of these polymers are a hypercrosslinked polystyreneproduced from crosslinked styrene copolymers by an extensivechloromethylation and a subsequent post-crosslinking by treating with aFriedel-Crafts catalyst in a swollen state. Yet other of these polymersare a hypercrosslinked polystyrene produced from crosslinked styrenecopolymers by an extensive additional post-crosslinking in a swollenstate with bifunctional crosslinking agents selected from the groupcomprising of monochlorodimethyl ether and p-xylilene dichloride

Some polymers useful in the practice of the invention are hydrophilicself wetting polymers that can be administered as dry powder containinghydrophilic functional groups such as, amines, hydroxyl, sulfonate, andcarboxyl groups.

Certain polymers useful in the invention are macroporous polymersprepared from the polymerizable monomers of styrene, divinylbenzene,ethylvinylbenzene, and the acrylate and methacrylate monomers such asthose listed below by manufacturer. Rohm and Haas Company, (now part ofDow Chemical Company): (i) macroporous polymeric sorbents such asAmberlite™ XAD-1, Amberlite™ XAD-2, Amberlite™ XAD-4, Amberlite™ XAD-7,Amberlite™ XAD-7HP, Amberlite™ XAD-8, Amberlite™ XAD-16, Amberlite™XAD-16 HP, Amberlite™ XAD-18, Amberlite™ XAD-200, Amberlite™ XAD-1180,Amberlite™ XAD-2000, Amberlite™ XAD-2005, Amberlite™ XAD-2010,Amberlite™ XAD-761, and Amberlite™ XE-305, and chromatographic gradesorbents such as Amberchrom™ CG 71,s,m,c, Amberchrom™ CG 161,s,m,c,Amberchrom™ CG 300,s,m,c, and Amberchrom™ CG 1000,s,m,c. Dow ChemicalCompany: Dowex® Optipore™ L-493, Dowex® Optipore™ V-493, Dowex®Optipore™ V-502, Dowex® Optipore™ L-285, Dowex® Optipore™ L-323, andDowex® Optipore™ V-503. Lanxess (formerly Bayer and Sybron): Lewatit®VPOC 1064 MD PH, Lewatit® VPOC 1163, Lewatit® OC EP 63, Lewatit® S6328A, Lewatit® OC 1066, and Lewatit® 60/150 MIBK. Mitsubishi ChemicalCorporation: Diaion® HP 10, Diaion® HP 20, Diaion® HP 21, Diaion® HP 30,Diaion® HP 40, Diaion® HP 50, Diaion® SP70, Diaion® SP 205, Diaion® SP206, Diaion® SP 207, Diaion® SP 700, Diaion® SP 800, Diaion® SP 825,Diaion® SP 850, Diaion® SP 875, Diaion® HP 1MG, Diaion® HP 2MG, Diaion®CHP 55A, Diaion® CHP 55Y, Diaion® CHP 20A, Diaion® CHP 20Y, Diaion® CHP2MGY, Diaion® CHP 20P, Diaion® HP 20SS, Diaion® SP 20SS, and Diaion® SP207SS. Purolite Company: Purosorb™ AP 250 and Purosorb™ AP 400.

As used herein, the term “sorbent” includes adsorbents and absorbents.

As used herein, the singular forms “a,” “an,” and “the” include theplural, and reference to a particular numerical value includes at leastthat particular value, unless the context clearly dictates otherwise.When a range of values is expressed, another embodiment includes fromthe one particular value and/or to the other particular value.Similarly, when values are expressed as approximations, by use of theantecedent “about,” it will be understood that the particular valueforms another embodiment. All ranges are inclusive and combinable.

The following examples are intended to be illustrative and not limiting.

Although not essential, a preferred embodiment would be a structure thatwas more selective for target compounds, such as an antibody orligand-coated beads or powders that are either non porous or porous. Inthe case of porous beads combined with an antibody, the antibody wouldaugment the broad removal properties of the porous polymer

EXPERIMENTAL Example 1 Sorbent Syntheses

The synthesis process consists of (1) preparing the aqueous phase, (2)preparing the organic phase, (3) carrying out the suspensionpolymerization, and (4) purifying the resulting porous polymeric sorbentproduct. The aqueous phase compositions are the same for all thepolymerizations. Table 1A lists the percentage composition of theaqueous phase and Table 1B gives the material charges typical for a five(5) liter-reactor polymerization run.

TABLE 1A Aqueous Phase Composition Wt. % Ultrapure Water 97.787Dispersing Agent: Polyvinylalcohol 0.290 Monosodium Phosphate 0.300Disodium Phosphate 1.000 Trisodium Phosphate 0.620 Sodium Nitrite 0.003

TABLE 1B Material Charges of a Typical Five (5) Liter-ReactorPolymerization Run Volume of Aqueous Phase 1750.00 ml Density of AqueousPhase 1.035 g/ml Weight of Aqueous Phase 1811.25 g Volumetric Ratio,Aqueous 1.05 Phase/Organic Phase Volume of Organic Phase 1665.0 mlDensity of Organic Phase 0.84093 g/ml Weight of Organic Phase, 1400.15 gExcluding Initiator Charge Total Reaction Volume 3415.0 ml TotalReaction Weight 3211.40 g

Upon preparation of the aqueous phase and the organic phase, the aqueousphase is poured into the five-liter reactor and heated to 65° C. withagitation. The pre-mixed organic phase including the initiator is pouredinto the reactor onto the aqueous phase with the stirring speed set atthe rpm for formation of the appropriate droplet size. The dispersion oforganic droplets is heated to the temperature selected for thepolymerization and is held at this temperature for the desired length oftime to complete the conversion of the monomers into the crosslinkedpolymer and, thereby, set the pore structure. Unreacted initiator isdestroyed by heating the bead slurry for two (2) hours at a temperaturewhere the initiator half-life is one hour or less. For the initiator,benzoyl peroxide, the unreacted initiator is destroyed by heating theslurry at 95° C. for two (2) hours.

The slurry is cooled, the mother liquor is siphoned from the beads andthe beads are washed five (5) times with ultrapure water. The beads arefreed of porogen and other organic compounds by a thermal cleaningtechnique. This process results in a clean, dry porous sorbent in theform of spherical, porous polymer beads.

TABLE 1C Components of Adsorbent Syntheses Porous Polymer Adsorbent 1Adsorbent 3 Adsorbent 4 Adsorbent 5 Identity Wt. %^(a) Adsorbent 2 Wt.%^(a) Wt. %^(a) Wt. %^(a) Divinylbenzene, 35.859 Adsorbent 2 26.16322.4127 22.4127 (DVB), Pure is a commercial Ethylvinylbenzene 20.141resin, 14.695 12.5883 12.5883 (EVB), Pure Amberlite Inerts 0.766XAD-16 ®, made 0.559 0.4790 0.4790 by Rohm and Haas Company Toluene19.234 27.263 64.521 54.841 Isooctane 24.00 31.319 0.00 9.680Polymerizable Monomers 56.00 40.8584 35.00 35.00 Porogen 44.00 59.141665.00 65.00 Benzoyl Peroxide 1.03 0.7447 2.00 4.00 (BPO), Pure; Wt. %Based Upon Polymerizable Monomer Content Polymerization, 75°/10 hrs80°/16 hrs 70°/24 hrs 65°/24 hrs ° C./time, hrs. 95°/2 hrs 95°/2 hrs^(a)Wt. % value is based upon the total weight of the organic phaseexcluding the initiator.

Example 2 Pore Structure Characterization

The pore structures of the sorbent polymer beds identified in TABLE 1Cwere analyzed with a Micromeritics ASAP 2010 instrument. The pore volumeis divided up into categories within pore size ranges for each of thefive sorbent polymers and these values are provided in TABLE 2. TheCapacity Pore Volume is that pore volume that is accessible to proteinsorption and consists of the pore volume in pores larger than 100 Ådiameter. The Effective Pore Volume is that pore volume that isselectively accessible to proteins smaller than 35,000 Daltons andconsists of pore diameters within the range of 100 to 250 Å diameter.The Oversized Pore Volume is the pore volume accessible to proteinslarger than 35,000 Daltons and consists of the pore volume in poreslarger than 250 Å diameter. The Undersize Pore Volume is the pore volumein pores smaller than 100 Å A diameter and is not accessible to proteinslarger than about 10,000 Daltons.

TABLE 2 Sorbent Sorbent Sorbent Sorbent Sorbent Porous Polymer Identity1 2 3 4 5 Pore Volume (cc/g) of pore 0.5512 1.509 1.711 0.67 0.89diameter in the range 50 Å to 3,000 Å Pore Volume (cc/g) of pore 0.0070.016 0.668 0.004 0.005 diameter in the range 500 Å to 3,000 Å Ratio ofPore Volume  84:1  95:1  3:1 183:1 170:1 between 50 Å to 3,000 Å indiameter To Pore Volume between 500 Å to 3,000 Å in diameter Pore Volume(cc/g) of pore 0.002 0.006 0.010 0.001 0.002 diameter in the range 1,000Å to 3,000 Å Ratio of Pore Volume 264:1 241:1 165:1 900:1 497:1 between50 Å to 3,000 Å in diameter To Pore Volume between 1,000 Å to 3,000 Å indiameter % Cytochrome-C, Sorbed 89% 97% 95%  57%  90% % Albumin Sorbed 4%  8% 13% 1.0% 1.8% Selectivity 24.05 11.94 7.27 57.1 50.06 PoreVolume (cc/g) of pore 0.573 1.214 1.555 0.264 0.67 diameter in the range100 to 3,000 Å Pore Volume (cc/g) of pore 0.211 0.711 1.345 0.012 0.02diameter in the range 200 to 3,000 Å Pore Volume (cc/g) of pore 0.0130.031 1.169 0.007 0.01 diameter in the range 300 to 3,000 Å

Example 3 Administration of a Sorbent to a Patient Exposed to an UnknownPathogen

A person is exposed to an unknown pathogen or toxin during an epidemicor bioterrorism attack. The mode of exposure is through one of manydifferent means such as food or water ingestion, aerosol inhalation orskin contact. The pathogen is one of many such as bacillus anthracis(anthrax), influenza virus, smallpox virus, Yersinia pestis (plague),Ebola or Marburg virus, Francisella tularensis (tularemia), hantavirus,cholera toxin, botulinum toxin, ricin toxin, Salmonella, Escherichiacoli such as E. coli 0157:H7, Shigella, Listeria, or others. Althoughthe organism is not yet identified, other people are rapidly becomingseriously ill with similar symptoms, including high fever, rigors,cough, severe fatigue, and diarrhea. The patient may receive standard ofcare therapy such as antivirals, antibiotics, antitoxins,immunoglobulins. Then, as a prophylactic measure or to treat a patientwho is developing symptoms of the infection and signs of inflammation(fever, chills, etc), the patient is orally administered milligram togram quantities of a slurry or suspension of the sorbent polymerparticulate or a tablet or capsule containing the sorbent polymer. Oncedistributed in the alimentary canal, particularly to the intestines, thesorbent captures, binds and sequesters inflammatory mediatorsexogenously introduced, or locally produced, or being introduced to thealimentary canal through physiologic fluids such as bile, before theseinflammatory mediators can induce further inflammation, or toxicity, orcause a degradation of the intestinal lining that can lead to worsenedinflammation, infection, endotoxemia and sepsis. By removing theseinflammatory mediators, the sorbent polymer reduces the triggers foradditional systemic inflammation in the patient, reducing systemicinflammatory mediator production, such as cytokines, thereby preventingor limiting the development of cytokine or other inflammatorymediator-induced cell death, organ damage, multi-organ failure andpotentially death. The sorbent is administered once or repeatedly overthe course of many hours to days to have a durable or persistent effect.The patient continues to receive antimicrobial therapy. The sorbent isthen excreted and eliminated from the patient's system, without beingmetabolized or degraded or otherwise retained in the body. The sorbentprevents the patient from developing severe sepsis, septic shock, orserious organ dysfunction.

Example 4 Administration of a Sorbent to a Patient with InflammatoryBowel Disease

A patient with inflammatory bowel disease develops an exacerbation withconstant severe diarrhea. The patient is administered standard of caretherapy which may include systemic steroids and parenteral anti-TNFtherapy. As part of the patients oral and rectal topicalanti-inflammatory regimen, the patient is administered an oralsuspension of the sorbent polymer as well as an enema containing thesorbent polymer. The sorbent binds, sequesters and removes locallyproduced inflammatory mediators, including cytokines, that are beingproduced enterally, and facilitates elimination of these mediatorsthrough excretion of the sorbent. This promotes healing of the intestineand a remission of the flare.

Example 5 Administration of a Sorbent to a Burn Patient

A burn patient suffers severe full thickness burns over 30% of his totalbody surface area as well as prolonged smoke and chemical inhalationlung injury. Despite debridement of the burns, the patient develops asevere systemic inflammatory response syndrome with the onset of acuterespiratory distress syndrome requiring mechanical ventilation. Thepatient is administered the standard of care, which involves mainlysupportive care measures. To counter the systemic inflammation that isleading rapidly to multi-organ failure, the patient is administered thesorbent polymer. Because the patient is unconscious, a feeding tube isused to deliver a slurry of the sorbent polymer to the alimentary canalto protect the intestinal lining from inflammatory mediators and tolimit systemic inflammation. The patient's blood can then be treatedextracorporeally by using a standard hemodialysis machine to pump theblood through a highly hemocompatible version of the sorbent, which iscapable of removing systemic inflammatory mediators that can lead to theproduction of more inflammatory mediators. Treatment with the sorbentoccurs over many days until the systemic inflammation subsides andinjured organs, including the lungs, begin to heal and recover.

Example 6 In Vitro Botulinum Neurotoxin Al Binding Study—ProtocolSummary

This study evaluates the ability of CytoSorbents' porous polymer beadsto bind and clear Botulinum Neurotoxin Type Al (BoNT/A1) from PhosphateBuffered Saline (PBS). The beads used in this study are representativeof the polymers described in this patent application. Divinylbenzenepolymer porous test beads and non-porous test beads were suspended insterile PBS (pH 7.4). A stock solution of BoNT/A1 in PBS was prepared ata concentration of 100 ug/mL (Tufts). Two (2) mL of a PBS solutioncontaining 100 ug/mL of BoNT/A1 (Tufts, Boston Mass.) was incubated insterile, wide-mouth 5 mL conical tubes (Stockwell #3206; 5 tubes/beadtype) containing either no beads, 100 uL dewetted non-porous beads, or100 uL dewetted porous beads (TDG-057-118-SS). The extrabead volume wasnominal in each case. 0.5 uL of the stock solution was removed andstored on ice (4° C.) for future analysis (T=0). Tubes were sealed andgently agitated for 60 minutes at room temperature. The beads wereallowed to settle and 0.5 mL samples of the supernatant were also placedon ice. Samples were evaluated for BoNT/A1 concentration via standardBCA (bicinchoninic acid) Protein Assay. The results demonstrate therelative impact of no bead, non-porous beads and porous beads in BoNT/A1removal. Porous beads reduced BoNT/A1 by 38.8% in this assay, comparedto an 8.6% removal by non-porous beads and a 3.9% increase in the nobead control. Results are presented in FIG. 1.

TABLE 2 BoNT/A1 removal by bead type. Time No (hr) Bead Non-PorousPorous 0 105.8 105.8 105.8 1 109.94 96.69 64.78

Example 7 Protocol Summary for Oral Administration of Porous PolymerBeads in a Cecal Ligation Puncture Sepsis Rat Model

This example investigates the therapeutic effect of a 25% oral dose, byvolume, of porous polymer beads on 19 week old Sprague-Dawley rats thathad undergone cecal ligation puncture (CLP) causing polymicrobialsepsis. The beads used in this study are representative of the polymersdescribed in this patent application.

Grouped in four cohorts of 8 animals (4 control and 4 treated), 32 ratswere acclimated for a minimum of three days prior to the start of thestudy; those that appear to be in obvious poor health were not be usedin the study. Animals were fasted for 16 hours before the surgicalprocedure was performed. Animals received subcutaneous injections ofBuprenorphine −0.1 mg/kg prior to surgery and then at 6-12 hourintervals post-surgery. All surgeries were performed using short actingisoflourane to minimize the deleterious effects of anesthesia oncardiovascular functions. The surgical procedure involved a 5 cm midlinelaparotomy beginning 2 cm below the sternum. The cecum was isolatedoutside of the abdominal cavity on sterile gauze to avoid blood vesseldamage. The cecum was then ligated with 2-0 Vicryl just below theileocecal valve, maintaining bowel continuity. The cecal contents werethen milked to one end of the cecum. The cecum was punctured 3 timeswith a 20-gauge needle, and then manipulated to extrude a single dropletof fecal material from each puncture site. The abdominal cavity was thenclosed in two layers, followed by fluid resuscitation and the animalreturned to the appropriate cage.

Animals were checked 2 hours post-surgery, prior to the end of the day,and first-thing the next morning. The rats were allowed food and waterad libitum. Approximately 20 hours post-surgery, living animals wereorally-dosed with 4 mL of either distilled water (N=8 for the controlgroup) or 25% polymer slurry in distilled water (N=14) using a steriledisposable gavage feeding tube. 12 hours later, the animals were orallydosed an additional 4 mL of 25% polymer slurry. Animals that died before20 hours post-CLP and before the first treatment dose were excluded fromthe study.

Detailed clinical observations were made and recorded on the day ofsurgery and on the day of dosing. Animals were observed for clinicalsigns once daily after dosing. Assessment included, but was not limitedto, activity, posture, respiration, hydration status, and overall bodycondition.

Euthanasia was performed via inhalation of isoflourane or carbon dioxidegas followed by cervical dislocation. Euthanized animals underwent agross necropsy and were discarded. Euthanasia was conducted inaccordance with accepted American Veterinary Medical Associationguidelines.

Prior to the first dosing, all animals demonstrated signs of a systemicinflammatory response, with cold paws, fur raising, hunched posture,inactivity and other clinical indicators of sepsis. Post-mortem grosspathology demonstrated erythema, edema and many surgical adhesions inthe peritoneal cavity amongst animals that died, while surviving animalshad markedly less pathology.

CRP Concentration Control vs. Treated (25% Slurry)

Effect of an Oral Dose of Porous Polymer on C-reactive protein (CRP)concentration over time. 16 Sprague Dawley rats were initiallyrandomized into each group (control and treated) and underwent cecalligation puncture (CLP) surgery in four separate cohorts of 4 controlanimals and 4 treatment animals. Animals that died in the pre-dosageperiod (<20 hours post-surgery) were excluded from these data. At 20 and32 hours after CLP, animals were gavaged with either 4 mL water(control) or 4mL of a 25% slurry of porous beads in water. The “0 Hours”time point on the graph is equivalent to 20 hours post-surgery. CRPConcentrations (ng/mL) were determined from plasma samples taken at timepoints of 0, 4, 8, 12, 16, and 28 hours beyond the “0 Hours” time pointin surviving animals. Results are presented in FIG. 2. The control groupresults are represented by the solid line (n=8) and the treated groupresults are represented by the dashed line (n=14). CRP is a generalmarker of systemic inflammation.

Percent of Animals Living vs. Time

Effect of an Oral Dose of Porous Polymer on Survival. 16 Sprague Dawleyrats were initially randomized into each group (control and treated) andunderwent cecal ligation puncture (CLP) surgery in four separate cohortsof 4 control animals and 4 treatment animals. Animals that died in thepre-dosage period (<20 hours post-surgery) were excluded from thesedata. At 20 and 32 hours after CLP, animals were gavaged with either 4mLwater (control) or 4mL of a 25% slurry of porous beads in water. The “0Hours” time point on the FIG. 3 is equivalent to 20 hours post-surgery.Survival time was observed from the “0 Hour” time point up to 27 hrs. At28 hours all animals were sacrificed. The Control group results arerepresented by the solid line (n=7) and the Treated group results arerepresented by the dashed line (n=14).

Example 8 Evaluation of Therapeutic Activity of an Orally AdministeredPorous Polymer Bead Slurry in the TNBS Colitis Model in Mice—ProtocolSummary

This example evaluates the therapeutic activity of an orallyadministered porous polymer bead slurry at two different dosesadministered twice a day in the TNBS (2,4,6-trinitrobenzenesulfonicacid) colitis model in mice. The beads used in this study arerepresentative of the polymers described in this patent application.

The health status of the animals used in this study was examined onarrival. Only animals in good health were acclimatized to laboratoryconditions and were used in the study. Animals were provided ad libitumwith a commercial sterile rodent diet and had free access to steriledrinking water that was supplied to each cage via polyethylene bottleswith stainless steel sipper tubes. Table 3 lists the experimental groupsthat comprised the study.

TABLE 3 Experimental Groups Group TNBS Group Size Test Item Dose VolumeRoute Dose Regime Induction 1 N = 16 Vehicle (DW) 0.150 ml 5 ml/kg POTwice a day TNBS in 40% DW* starting on ethanol 2 N = 16 TDG-057-145,0.150 ml* 5 ml/kg PO study day 0, on study day 0 Modified, (50% 10 hourspost Batch 1, compound TNBS induction −106/+45 suspended and through the50% in water) end of the study.* 3 N = 4  Sham 0.150 ml 5 ml/kg PO Twicea day 40% ethanol DW* starting on on study day 0 study day 0. *NOTE:Immediately after Group 2 was dosed with 0.150 ml of TI, an additional0.150 ml of water was drawn into the same syringe used to gavage theanimals and administered. This 2^(nd) step washed down any residualcompound in the syringe to assure maximum dose delivery. Groups 1 and 3were dosed with 0.150 ml of DW and followed by an additional 0.150 ml ofDW which was drawn into the same syringe used to gavage the animals andadministered.

Animals from Groups 1-3 were induced with 80 μL (0.8 mL) TNBS dissolvedin 80% ethanol at 1:1 ratio (final ethanol concentration=40%) into thecolon, 4 cm proximal to the anus through the rectum. The animals werekept in a vertical position for 60 seconds before being returned totheir cages. Animals from group 3 were only given 40% ethanol as anegative control of the induced groups.

All TI dosing solutions were applied twice daily on each of the repeateddosing sessions starting on Study Day 0, 10 hours post-TNBS inductionand through the end of the study (Day 7). For the control animals ingroups 1 and 3, only water was be gavaged in place of TI dosing.

Prior to gavage feeding, the following procedures were performed:

1) The samples were rocked/swirled to evenly suspend the polymer whileavoiding entraining air.2) The syringes were primed once with water to fill all void spaces inthe syringe needle and barrel and water was expelled to remove airbubbles. The mouse was gavaged IMMEDIATELY to prevent Test Itemsettling.3) Between animals, samples were gently mixed to ensure a homogeneoussuspension. If there was a delay in gavaging the animal after drawing upthe polymer slurry, the contents in the syringe were mixed by invertingthe syringe several times to ensure homogeneity.4) After the animal was dosed with 0.150 mL of TI, an additional 0.150mL of water was drawn into the same syringe to gavage the animal. Thisstep assured the maximum dose delivery.5) When not in use, the polymer slurries were stored in a refrigerator.

Throughout the entire 8-day duration of the study, careful clinicalexaminations of the animals were carried out in a blinded fashion andrecorded once daily. Observations included changes in skin, fur, eyes,mucous membranes, occurrence of secretions and excretions (e.g.diarrhea) and autonomic activity (e.g. lacrimation, salivation,piloerection, pupil size, unusual respiratory pattern). Changes in gait,posture and response to handling, as well as the presence of unusualbehavior, tremors, convulsions, sleep and coma were also noted.

Individual body weights of each animal were measured on Study Day 0(Study Commencement) and on a daily basis thereafter through studytermination (Study Day 7).

NOTE: If an animal died or was euthanized during the study, its lastweight was kept constant for the remaining days. This method allowedsome representation of this animal throughout the study.

Body weight, stool consistency and bleeding per rectum, as well as anyabnormalities observed in fur coat and abdomen were recorded daily.Total clinical score was presented as a SUM of all the parameters listedin the Table 4.

TABLE 4 Total Clinical Scores Score Weight Loss % None 0  1-5 1  5-10 210-15 3 >15 4 Death 11  Stool Consistency Normal 0 Redness, swelling ofthe anus 1 Loose stool 2 Diarrhea 3 Diarrhea + Blood 4 — — Fur Coat*None 0 Fur 1 Fur 1 Fur 1 Fur 1 — — Abdomen* None 0 Abdomen 1 Abdomen 1Abdomen 1 Abdomen 1 — — *NOTE: Abnormalities in fur coat and abdomenincluded but were not limited to: shaggy fur and swollen abdomen.

All living animals were euthanized at the end of the study.

Animals found in a moribund condition and/or animals showing severe painand enduring signs of severe distress were humanely euthanized. Inaddition, and unless decided otherwise in the course of the study,animals that showed a decrease of body weight greater than 20% frominitial body weight determination were humanely euthanized in accordancewith animal regulations. All such events were recorded in the finalreport.

Results from the blinded evaluation are presented in FIG. 4.

What is claimed:
 1. A method of treating systemic, regional, or localinflammation from a patient suffering inflammation, said methodcomprising administration of a therapeutically effective dose of asorbent for an inflammatory mediator in said patient; wherein thesorbent is a biocompatible polymer that comprises a cross-linked, porousstyrene or divinylbenzene copolymer; the biocompatible polymer having apore structure that the total pore volume of pore size in the range of50 Å to 3000 Å is greater than 0.5 cc/g to 3.0 cc/g dry polymer; whereinsaid inflammatory mediator results from inflammatory bowel disease,burns to said patient, exposure of said patient to neurotoxins orsepsis.
 2. The method of claim 1, wherein said inflammatory mediator ispresent in physiological fluid or a carrier fluid in the patient.
 3. Themethod of claim 2, wherein said physiological fluids is nasopharyngeal,oral, esophageal, gastric, pancreatic, hepatic, pleural, pericardial,peritoneal, intestinal, prostatic, seminal, vaginal secretions, tears,saliva, mucus, bile, blood, lymph, plasma, serum, synovial fluid,cerebrospinal fluid, urine, and interstitial, intracellular, orextracellular fluid.
 4. The method of claim 1, wherein said dose ofsorbent is administered orally, via a feeding tube, peritoneally, orrectally.
 5. The method of claim 1, wherein said biocompatible polymercomprises particles having a diameter in the range for 0.1 micron metersto 2 centimeters.
 6. The method of claim 5, wherein said biocompatiblepolymer is in the form of powder, beads or other regular or irregularlyshaped particulate.
 7. The method of claim 1, wherein: the Ratio of PoreVolume between 50 Å to 3,000 Å in diameter To Pore Volume between 500 Åto 3,000 Å in diameter of the said biocompatible polymer is smaller than200:1; and wherein the Ratio of Pore Volume between 50 Å to 3,000 Å indiameter To Pore Volume between 1,000 Å to 3,000 Å in diameter of saidbiocompatible polymer is greater than 20:1.
 8. The method of claim 1,wherein said biocompatible polymer is a coated polymer comprising atleast one crosslinking agent and at least one dispersing agent; saiddispersing agents such as hydroxyethyl cellulose, hydroxypopylcellulose, poly(hydroxyethyl methacrylate), poly(hydroxyethyl acrylate),poly(hydroxypropyl methacrylate), poly(hydroxypropyl acrylate),poly(dimethylaminoethyl methacrylate), poly(dimethylaminoethylacrylate), poly(diethylamimoethyl methacrylate), poly(diethylaminoethylacrylate), poly(vinyl alcohol), poly(N-vinylpyrrolidinone), salts ofpoly(methacrylic acid), and salts of poly(acrylic acid) and mixturesthereof; said crosslinking agent selected from a group consisting ofdivinylbenzene, trivinylbenzene, divinylnaphthalene,trivinylcyclohexane, divinylsulfone, trimethylolpropane trimethacrylate,trimethylolpropane dimethacrylate, trimethylolpropane triacrylate,trimethylolpropane diacrylate, pentaerythrital dimethacrylates,pentaerythrital trimethacrylates, pentaerythrital, tetramethacrylates,pentaerythritol diacrylates, pentaerythritol triiacrylates,pentaerythritol tetraacrylates, dipentaerythritol dimethacrylates,dipentaerythritol trimethacrylates, dipentaerythritoltetramethacrylates, dipentaerythritol diacrylates, dipentaerythritoltriacrylates, dipentaerythritol tetraacrylates, divinylformamide andmixtures thereof; said polymer being developed simultaneously with theformation of the coating, wherein said dispersing agent is chemicallybound to said surface of said biocompatible polymer.
 9. The method ofclaim 8, wherein said biocompatible polymer comprises residues from oneor more monomers selected from divnylbenzene and ethylvinylbezene,styrene, ethylstyrene, acrylonitrile, butyl methacrylate, octylmethacrylate, butyl acrylate, octyl acrylate, cetyl methacrylate, cetylacrylate, ethyl methacrylate, ethyl acrylate, vinyltoluene,vinylnaphthalene, vinylbenzyl alcohol, vinylformamide, methylmethacrylate, methyl acrylate, trivinylbenzene, divinylnaphthalene,trivinylcyclohexane, divinylsulfone, trimethylolpropane trimethacrylate,trimethylolpropane dimethacrylate, trimethylolpropane triacrylate,trimethylolpropane diacrylate, pentaerythritol dimethacrylate,pentaerythritol trimethacrylate, pentaerythritol tetramethacrylate,pentaerythritol diacrylate, pentaerythritol triiacrylate,pentaerythritol tetraacrylate, dipentaerythritol dimethacrylate,dipentaerythritol trimethacrylate, dipentaerythritol tetramethacrylate,dipentaerythritol diacrylate, dipentaerythritol triacrylate,dipentaerythritol tetraacrylate, divinylformamide and mixtures thereof.10. The method of claim 1, wherein the sorbent is supplied as slurry, orsuspension, or dry powder or other dry particulate capable of beingwetted.
 11. A method of treating systemic, regional, or localinflammation from a patient suffering inflammation, said methodcomprising administration of a therapeutically effective dose of asorbent for an inflammatory mediator in said patient; wherein thesorbent is a biocompatible polymer that comprises a cross-linked, porousstyrene or divinylbenzene copolymer; the biocompatible polymer having apore structure that the total pore volume of pore size in the range of50 Å to 3000 Å is greater than 0.5 cc/g to 3.0 cc/g dry polymer; whereinsaid inflammatory mediator comprises is one or more of enzymes,prostaglandins, eicosanoids, leukotrienes, kinins, inflammatory mediatorassociated with anaphylaxis, coagulation factors, endotoxins,enterotoxins, lipopolysaccharide, cell fragments, bile salts, fattyacids, phospholipids, reactive oxygen species, oxygen radicals,surfactants, interferon and immunomodulatory antibodies, biologics ordrugs, including those administered exogenously .
 12. The method ofclaim 11, wherein said inflammatory mediator is present in physiologicalfluid or a carrier fluid in the patient.
 13. The method of claim 12,wherein said physiological fluids is nasopharyngeal, oral, esophageal,gastric, pancreatic, hepatic, pleural, pericardial, peritoneal,intestinal, prostatic, seminal, vaginal secretions, tears, saliva,mucus, bile, blood, lymph, plasma, serum, synovial fluid, cerebrospinalfluid, urine, and interstitial, intracellular, or extracellular fluid.14. The method of claim 11, wherein said dose of sorbent is administeredorally, via a feeding tube, peritoneally, or rectally.
 15. The method ofclaim 11, wherein said biocompatible polymer comprises particles havinga diameter in the range for 0.1 micron meters to 2 centimeters.
 16. Themethod of claim 11, wherein said biocompatible polymer is in the form ofpowder, beads or other regular or irregularly shaped particulate. 17.The method of claim 11, wherein the sorbent is supplied as slurry, orsuspension, or dry powder or other dry particulate capable of beingwetted.
 18. The method of claim 11, wherein: said biocompatible polymerhas a pore structure that the total pore volume of pore size in therange of 50 Å to 3000 Å is greater than 0.5 cc/g to 3.0 cc/g drypolymer; wherein the Ratio of Pore Volume between 50 Å to 3,000 Å indiameter To Pore Volume between 500 Å to 3,000 Å in diameter of the saidbiocompatible polymer is smaller than 200:1; and wherein the Ratio ofPore Volume between 50 Å to 3,000 Å in diameter To Pore Volume between1,000 Å to 3,000 Å in diameter of said biocompatible polymer is greaterthan 20:1.
 19. The method of claim 11, wherein said biocompatiblepolymer is a coated polymer comprising at least one crosslinking agentand at least one dispersing agent; said dispersing agents such ashydroxyethyl cellulose, hydroxypopyl cellulose, poly(hydroxyethylmethacrylate), poly(hydroxyethyl acrylate), poly(hydroxypropylmethacrylate), poly(hydroxypropyl acrylate), poly(dimethylaminoethylmethacrylate), poly(dimethylaminoethyl acrylate), poly(diethylamimoethylmethacrylate), poly(diethylaminoethyl acrylate), poly(vinyl alcohol),poly(N-vinylpyrrolidinone), salts of poly(methacrylic acid), and saltsof poly(acrylic acid) and mixtures thereof; said crosslinking agentselected from a group consisting of divinylbenzene, trivinylbenzene,divinylnaphthalene, trivinylcyclohexane, divinylsulfone,trimethylolpropane trimethacrylate, trimethylolpropane dimethacrylate,trimethylolpropane triacrylate, trimethylolpropane diacrylate,pentaerythrital dimethacrylates, pentaerythrital trimethacrylates,pentaerythrital, tetramethacrylates, pentaerythritol diacrylates,pentaerythritol triiacrylates, pentaerythritol tetraacrylates,dipentaerythritol dimethacrylates, dipentaerythritol trimethacrylates,dipentaerythritol tetramethacrylates, dipentaerythritol diacrylates,dipentaerythritol triacrylates, dipentaerythritol tetraacrylates,divinylformamide and mixtures thereof; said polymer being developedsimultaneously with the formation of the coating, wherein saiddispersing agent is chemically bound to said surface of saidbiocompatible polymer.
 20. The method of claim 19, wherein saidbiocompatible polymer comprises residues from one or more monomersselected from divnylbenzene and ethylvinylbezene, styrene, ethylstyrene,acrylonitrile, butyl methacrylate, octyl methacrylate, butyl acrylate,octyl acrylate, cetyl methacrylate, cetyl acrylate, ethyl methacrylate,ethyl acrylate, vinyltoluene, vinylnaphthalene, vinylbenzyl alcohol,vinylformamide, methyl methacrylate, methyl acrylate, trivinylbenzene,divinylnaphthalene, trivinylcyclohexane, divinylsulfone,trimethylolpropane trimethacrylate, trimethylolpropane dimethacrylate,trimethylolpropane triacrylate, trimethylolpropane diacrylate,pentaerythritol dimethacrylate, pentaerythritol trimethacrylate,pentaerythritol tetramethacrylate, pentaerythritol diacrylate,pentaerythritol triiacrylate, pentaerythritol tetraacrylate,dipentaerythritol dimethacrylate, dipentaerythritol trimethacrylate,dipentaerythritol tetramethacrylate, dipentaerythritol diacrylate,dipentaerythritol triacrylate, dipentaerythritol tetraacrylate,divinylformamide and mixtures thereof.